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Human 2'-deoxynucleoside 5'-phosphate N-hydrolase 1 : the catalytic roles of Tyr24 and Asp80
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dc.contributor.author | Carberry, Anna Ellen | |
dc.contributor.author | Devi, Suneeta | |
dc.contributor.author | Harrison, David James | |
dc.contributor.author | da Silva, R.G. | |
dc.date.accessioned | 2024-03-01T14:30:01Z | |
dc.date.available | 2024-03-01T14:30:01Z | |
dc.date.issued | 2024-04-02 | |
dc.identifier | 299324030 | |
dc.identifier | 56c6b2bc-aea5-4837-8a33-812ed3700986 | |
dc.identifier | 85186397524 | |
dc.identifier.citation | Carberry , A E , Devi , S , Harrison , D J & da Silva , R G 2024 , ' Human 2'-deoxynucleoside 5'-phosphate N-hydrolase 1 : the catalytic roles of Tyr24 and Asp80 ' , ChemBioChem , vol. 25 , no. 7 , e202400047 . https://doi.org/10.1002/cbic.202400047 | en |
dc.identifier.issn | 1439-4227 | |
dc.identifier.other | ORCID: /0000-0001-9041-9988/work/154531736 | |
dc.identifier.other | ORCID: /0000-0002-1308-8190/work/154532445 | |
dc.identifier.uri | https://hdl.handle.net/10023/29399 | |
dc.description.abstract | The human enzyme 2′-deoxynucleoside 5′-phosphate N-hydrolase 1 (HsDNPH1) catalyses the hydrolysis of 5-hydroxymethyl-2′-deoxyuridine 5′-phosphate to generate 5-hydroxymethyluracil and 2-deoxyribose-5-phosphate via a covalent 5-phospho-2-deoxyribosylated enzyme intermediate. HsDNPH1 is a promising target for inhibitor development towards anticancer drugs. Here, site-directed mutagenesis of conserved active-site residues, followed by HPLC analysis of the reaction and steady-state kinetics are employed to reveal the importance of each of these residues in catalysis, and the reaction pH-dependence is perturbed by each mutation. Solvent deuterium isotope effects indicate no rate-limiting proton transfers. Crystal structures of D80N-HsDNPH1 in unliganded and substrate-bound states, and of unliganded D80A- and Y24F-HsDNPH1 offer atomic level insights into substrate binding and catalysis. The results reveal a network of hydrogen bonds involving the substrate and the E104-Y24-D80 catalytic triad and are consistent with a proposed mechanism whereby D80 is important for substrate positioning, for helping modulate E104 nucleophilicity, and as the general acid in the first half-reaction. Y24 positions E104 for catalysis and prevents a catalytically disruptive close contact between E104 and D80. | |
dc.format.extent | 10 | |
dc.format.extent | 2055543 | |
dc.language.iso | eng | |
dc.relation.ispartof | ChemBioChem | en |
dc.subject | DNPH1 | en |
dc.subject | pH-Rate profiles | en |
dc.subject | Biocatalysis | en |
dc.subject | Cancer | en |
dc.subject | Isotope effects | en |
dc.subject | QD Chemistry | en |
dc.subject | DAS | en |
dc.subject.lcc | QD | en |
dc.title | Human 2'-deoxynucleoside 5'-phosphate N-hydrolase 1 : the catalytic roles of Tyr24 and Asp80 | en |
dc.type | Journal article | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Sir James Mackenzie Institute for Early Diagnosis | en |
dc.contributor.institution | University of St Andrews. Cellular Medicine Division | en |
dc.contributor.institution | University of St Andrews. School of Medicine | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | 10.1002/cbic.202400047 | |
dc.description.status | Peer reviewed | en |
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